Abstract
Liquid inoculum is widely adopted in the mushroom industry, yet preparing Tremella fuciformis liquid inoculum remains challenging due to its complex microbial community and dimorphic growth. This study aimed to establish a reliable protocol for T. fuciformis liquid inoculum and assess its practical application. Initially, liquefied spawn was produced by liquefying solid spawn. The application of standard liquefied spawn increased fruiting body yield by 8.2% (502.4 g/kg dry substrate) compared to solid spawn, but exhibited substantial batch-to-batch variation due to unstable microbial communities and low Tremellomycetes abundance. To address these limitations, liquid spawn was developed via pre-culture of pure T. fuciformis and Annulohypoxylon stygium mycelia. Cultivation tests demonstrated significantly enhanced performance with 608.2 g of fruiting bodies, which represented 11.1% improvement compared to solid spawn. Moreover, consistent yields could be observed across multiple batches. This stability was attributed to stable microbial community structure and the dominance of Tremellomycetes (abundance > 50%) in the fungal community. These results confirm the cultivation performance of T. fuciformis liquid spawn, highlighting its potential as an effective alternative to solid spawn for T. fuciformis industrial production.